Regeneration Potential of Jellyfish: Cellular Mechanisms and Molecular Insights.

Medusozoans, the Cnidarian subphylum, have multiple life stages including sessile polyps and free-swimming medusae or jellyfish, which are typically bell-shaped gelatinous zooplanktons that exhibit diverse morphologies. Despite having a relatively complex body structure with well-developed muscles and nervous systems, the adult medusa stage maintains a high regenerative ability that enables organ regeneration as well as whole body reconstitution from the part of the body. This remarkable regeneration potential of jellyfish has long been acknowledged in different species; however, recent studies have begun dissecting the exact processes underpinning regeneration events. In this article, we introduce the current understanding of regeneration mechanisms in medusae, particularly focusing on cellular behaviors during regeneration such as wound healing, blastema formation by stem/progenitor cells or cell fate plasticity, and the organism-level patterning that restores radial symmetry. We also discuss putative molecular mechanisms involved in regeneration processes and introduce a variety of novel model jellyfish species in the effort to understand common principles and diverse mechanisms underlying the regeneration of complex organs and the entire body.

Gastrulation and germ layer formation in the sea anemone Nematostella vectensis and other cnidarians.

Among the basally branching metazoans, cnidarians display well-defined gastrulation processes leading to a diploblastic body plan, consisting of an endodermal and an ectodermal cell layer. As the outgroup to all Bilateria, cnidarians are an interesting group to investigate ancestral developmental mechanisms. Interestingly, all known gastrulation mechanisms known in Bilateria are already found in different species of Cnidaria. Here I review the morphogenetic processes found in different Cnidaria and focus on the investigation of the cellular and molecular mechanisms in the sea anemone Nematostella vectensis, which has been a major model organism among cnidarians for evolutionary developmental biology. Many of the genes involved in germ layer specification and morphogenetic processes in Bilateria are also found active during gastrulation of Nematostella and other cnidarians, suggesting an ancestral role of this process. The molecular analyses indicate a tight link between gastrulation and axis patterning processes by Wnt and FGF signaling. Interestingly, the endodermal layer displays many features of the mesodermal layer in Bilateria, while the pharyngeal ectoderm has an endodermal expression profile. Comparative analyses as well as experimental studies using embryonic aggregates suggest that minor differences in the gene regulatory networks allow the embryo to transition relatively easily from one mode of gastrulation to another.

Relative roles of biological and physical processes influencing coral recruitment during the lag phase of reef community recovery.

Following disturbances, corals recolonize space through the process of recruitment consisting of the three phases of propagule supply, settlement, and post-settlement survival. Yet, each phase is influenced by biophysical factors, leading to recruitment success variability through space. To resolve the relative contributions of biophysical factors on coral recruitment, the recovery of a 150 km long coral reefs in Palau was investigated after severe typhoon disturbances. Overall, we found that benthic organisms had a relatively weak interactive influence on larval settlement rates at the scale of individual tiles, with negative effects mainly exerted from high wave exposure for Acropora corals. In contrast, juvenile coral densities were well predicted by biophysical drivers, through both direct and indirect pathways. High densities of Acropora and Poritidae juveniles were directly explained by the availability of substrata free from space competitors. Juvenile Montipora were found in higher densities where coralline algae coverage was high, which occurred at reefs with high wave exposure, while high densities of juvenile Pocilloporidae occurred on structurally complex reefs with high biomass of bioeroder fish. Our findings demonstrate that strengths of biophysical interactions were taxon-specific and had cascading effects on coral recruitment, which need consideration for predicting reef recovery and conservation strategies.

Chromatin dynamics enable transcriptional rhythms in the cnidarian Nematostella vectensis.

In animals, circadian rhythms are driven by oscillations in transcription, translation, and proteasomal degradation of highly conserved genes, resulting in diel cycles in the expression of numerous clock-regulated genes. Transcription is largely regulated through the binding of transcription factors to cis-regulatory elements within accessible regions of the chromatin. Chromatin remodeling is linked to circadian regulation in mammals, but it is unknown whether cycles in chromatin accessibility are a general feature of clock-regulated genes throughout evolution. To assess this, we applied an ATAC-seq approach using Nematostella vectensis, grown under two separate light regimes (light:dark (LD) and constant darkness (DD)). Based on previously identified N. vectensis circadian genes, our results show the coupling of chromatin accessibility and circadian transcription rhythmicity under LD conditions. Out of 180 known circadian genes, we were able to list 139 gene promoters that were highly accessible compared to common promoters. Furthermore, under LD conditions, we identified 259 active enhancers as opposed to 333 active enhancers under DD conditions, with 171 enhancers shared between the two treatments. The development of a highly reproducible ATAC-seq protocol integrated with published RNA-seq and ChIP-seq databases revealed the enrichment of transcription factor binding sites (such as C/EBP, homeobox, and MYB), which have not been previously associated with circadian signaling in cnidarians. These results provide new insight into the regulation of cnidarian circadian machinery. Broadly speaking, this supports the notion that the association between chromatin remodeling and circadian regulation arose early in animal evolution as reflected in this non-bilaterian lineage.

Regionalized nervous system in Hydra and the mechanism of its development.

The last common ancestor of Bilateria and Cnidaria is considered to develop a nervous system over 500 million years ago. Despite the long course of evolution, many of the neuron-related genes, which are active in Bilateria, are also found in the cnidarian Hydra. Thus, Hydra is a good model to study the putative primitive nervous system in the last common ancestor that had the great potential to evolve to a more advanced one. Regionalization of the nervous system is one of the advanced features of bilaterian nervous system. Although a regionalized nervous system is already known to be present in Hydra, its developmental mechanisms are poorly understood. In this study we show how it is formed and maintained, focusing on the neuropeptide Hym-176 gene and its paralogs. First, we demonstrate that four axially localized neuron subsets that express different combination of the neuropeptide Hym-176 gene and its paralogs cover almost an entire body, forming a regionalized nervous system in Hydra. Second, we show that positional information governed by the Wnt signaling pathway plays a key role in determining the regional specificity of the neuron subsets as is the case in bilaterians. Finally, we demonstrated two basic mechanisms, regionally restricted new differentiation and phenotypic conversion, both of which are in part conserved in bilaterians, are involved in maintaining boundaries between the neuron subsets. Therefore, this study is the first comprehensive analysis of the anatomy and developmental regulation of the divergently evolved and axially regionalized peptidergic nervous system in Hydra, implicating an ancestral origin of neural regionalization.

Epithelial folding in the morphogenesis of the colonial marine hydrozoan, Dynamena pumila.

Epithelial folding (EF) is a fundamental morphogenetic process that can be observed in the development of many organisms ranging from metazoans to green algae. Being early branching metazoans, cnidarians represent the best models to study evolutionarily conserved morphogenetic processes, including EF. Hydrozoa is the most evolutionary advanced group of the phylum Cnidaria. All colonial hydrozoans grow continuously, changing the shape of their colonies and spreading over the substrate with the help of elongating stolons. Owing to high diversity of colony architecture, they are ideal objects for comparative and evolutionary morphology. In the hydrozoan Dynamena pumila, the growth of the colony proceeds via a variety of morphogenetic processes. Our work is focused on the formation of the anchoring disc of the stolon, which is accompanied by inward-folding morphogenesis of the ectodermal layer. Successive stages of anchoring disc development were described with light, confocal transmission electron microscopy. We have shown that EF in Dynamena is associated with accumulation of F-actin in the constricting apical domains of forming bottle cells located at the bottom of the emerging fold. In addition, the nuclei of these cells are displaced to the basal domains. Taken together, these features may indicate that EF in Dynamena proceeds as an active invagination, although this process has never been described in the development of hydrozoans. Apparently, development of the anchoring disc can be viewed as a reliable and versatile model system for studying the cell-shape-change-driven epithelial sheet morphogenesis, which can be easily observed and analysed.

Symbiont Identity Influences Patterns of Symbiosis Establishment, Host Growth, and Asexual Reproduction in a Model Cnidarian-Dinoflagellate Symbiosis.

The genus Symbiodinium is physiologically diverse and so may differentially influence symbiosis establishment and function. To explore this, we inoculated aposymbiotic individuals of the sea anemone Exaiptasia pallida (commonly referred to as "Aiptasia"), a model for coral symbiosis, with one of five Symbiodinium species or types (S. microadriaticum, S. minutum, phylotype C3, S. trenchii, or S. voratum). The spatial pattern of colonization was monitored over time via confocal microscopy, and various physiological parameters were measured to assess symbiosis functionality. Anemones rapidly formed a symbiosis with the homologous symbiont, S. minutum, but struggled or failed to form a long-lasting symbiosis with Symbiodinium C3 or S. voratum, respectively. Symbiodinium microadriaticum and S. trenchii were successful but reached their peak density two weeks after S. minutum. The spatial pattern of colonization was identical for all Symbiodinium taxa that were ultimately successful, starting in the oral disk and progressing to the tentacles, before invading the column and, finally, the pedal disk. In all cases, proliferation through the anemone's tentacles was patchy, suggesting that symbionts were being expelled into the gastrovascular cavity and re-phagocytosed by the host. However, the timing of these various spatial events differed between the different Symbiodinium taxa. Furthermore, S. microadriaticum and S. trenchii were less beneficial to the host, as indicated by lower rates of photosynthesis, anemone growth, and pedal laceration. This study enhances our understanding of the link between symbiont identity and the performance of the overall symbiosis, which is important for understanding the potential establishment and persistence of novel host-symbiont pairings. Importantly, we also provide a baseline for further studies on this topic with the globally adopted "Aiptasia" model system.

Back to the Basics: Cnidarians Start to Fire.

The nervous systems of cnidarians, pre-bilaterian animals that diverged close to the base of the metazoan radiation, are structurally simple and thus have great potential to reveal fundamental principles of neural circuits. Unfortunately, cnidarians have thus far been relatively intractable to electrophysiological and genetic techniques and consequently have been largely passed over by neurobiologists. However, recent advances in molecular and imaging methods are fueling a renaissance of interest in and research into cnidarians nervous systems. Here, we review current knowledge on the nervous systems of cnidarian species and propose that researchers should seize this opportunity and undertake the study of members of this phylum as strategic experimental systems with great basic and translational relevance for neuroscience.

The cellular and molecular basis of cnidarian neurogenesis.

Neurogenesis initiates during early development and it continues through later developmental stages and in adult animals to enable expansion, remodeling, and homeostasis of the nervous system. The generation of nerve cells has been analyzed in detail in few bilaterian model organisms, leaving open many questions about the evolution of this process. As the sister group to bilaterians, cnidarians occupy an informative phylogenetic position to address the early evolution of cellular and molecular aspects of neurogenesis and to understand common principles of neural development. Here we review studies in several cnidarian model systems that have revealed significant similarities and interesting differences compared to neurogenesis in bilaterian species, and between different cnidarian taxa. Cnidarian neurogenesis is currently best understood in the sea anemone Nematostella vectensis, where it includes epithelial neural progenitor cells that express transcription factors of the soxB and atonal families. Notch signaling regulates the number of these neural progenitor cells, achaete-scute and dmrt genes are required for their further development and Wnt and BMP signaling appear to be involved in the patterning of the nervous system. In contrast to many vertebrates and Drosophila, cnidarians have a high capacity to generate neurons throughout their lifetime and during regeneration. Utilizing this feature of cnidarian biology will likely allow gaining new insights into the similarities and differences of embryonic and regenerative neurogenesis. The use of different cnidarian model systems and their expanding experimental toolkits will thus continue to provide a better understanding of evolutionary and developmental aspects of nervous system formation. WIREs Dev Biol 2017, 6:e257. doi: 10.1002/wdev.257 For further resources related to this article, please visit the WIREs website.

[The gastrulation in Cnidaria: A key to understanding phylogeny or the chaos of secondary modifications?].

The data revealed by comparative embryology of the basal (diploblastic) metazoans is traditionally considered a valuable potential source of information on the origin and early evolution of the animal kingdom and its major clades. Special attention is paid to the fundamental morphogenetic process of gastrulation during which the cells of the early embryo differentiate into the germ layers and the primary body plan is formed. Comparative analysis of gastrulation in different cnidarian taxa reveals high level of intergroup, intragroup, and individual variation. With few exceptions, there is no robust correlation between the type of gastrulation and the taxon. Current data do not support the idea that morphogenetic processes underlying cnidarian gastrulation can be divided into several distinct types. Rather, there is a continuum of equifinal ontogenetic trajectories. In cnidarians, the mode of gastrulation apparently depends less on the macroevolutionary history of the species than on various evolutionary plastic features, such as the oocyte size, the amount of yolk, the number of cells at the blastula (or morula) stage, the presence of phototrophic symbionts, or the ecology of the larva. Thus, in cnidarians, morphogenetic basis of gastrulation contains only a very weak phylogenetic signal and can have only limited application in phylogenetic reconstructions. On the other hand, comparative studies of the ontogeny of the basal metazoans shed light on the general rules of the evolution of morphogenetic processes that is crucial for understanding the early history of the animal kingdom.

Meeting report on "Animal Evolution: New Perspectives From Early Emerging Metazoans", Tutzing, September 14-17, 2015.

This recent meeting covered non-bilaterian (e.g., cnidarians, ctenophores, and sponges) animals broadly, but with emphasis in four areas: 1) New genomic resources and tools for functional studies, 2) advances in developmental and regeneration studies, 3) the evolution and function of nervous systems, 4) symbiosis and the holobiome.

Composition and biological activities of the aqueous extracts of three scleractinian corals from the Mexican Caribbean: Pseudodiploria strigosa, Porites astreoides and Siderastrea siderea

Background: Scleractinian corals (stony corals) are the most abundant reef-forming cnidarians found in coral reefs throughout the world. Despite their abundance and ecological importance, information about the diversity of their toxins and their biological activities is very scarce. In this study, the chemical composition and the biological activities of the aqueous extracts of Pseudodiploria strigosa, Porites astreoides and Siderastrea siderea, three scleractinian corals from the Mexican Caribbean, have been assessed for the first time. Methods: Toxicity of the extracts was assessed in crickets; the presence of cytolysins was detected by the hemolysis assay; the vasoconstrictor activity was determined by the isolated rat aortic ring assay; the nociceptive activity was evaluated by the formalin test. The presence of phospholipases A2 (PLA2), serine proteases, and hyaluronidases was determined by enzymatic methods. Low-molecular-weight fractions were obtained by gel filtration chromatography and ultrafiltration. Results: Extracts from the three species were toxic to crickets, induced hemolysis in human and rat erythrocytes, produced vasoconstriction on isolated rat aortic rings, and presented phospholipase A2 and serine-protease activity. Despite the fact that these corals are not considered to be harmless to humans, the extracts generated significant nociceptive responses. The matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry analysis of the low-molecular-weight fractions revealed the presence of peptides within a mass range of 3000 to 6000 Da. These fractions were toxic to crickets and two of them induced a transitory vasoconstrictor effect on isolated rat aortic rings. Conclusion: This study suggests that scleractinian corals produce low-molecular-weight peptides that are lethal to crickets and induce vasoconstriction.(AU)

Mass Mortality Events in the NW Adriatic Sea: Phase Shift from Slow- to Fast-Growing Organisms.

Massive outbreaks are increasing all over the world, which are likely related to climate change. The North Adriatic Sea, a sub-basin of the Mediterranean Sea, is a shallow semi-closed sea receiving high nutrients inputs from important rivers. These inputs sustain the highest productive basin of the Mediterranean Sea. Moreover, this area shows a high number of endemisms probably due to the high diversity of environmental conditions and the conspicuous food availability. Here, we documented two massive mortalities (2009 and 2011) and the pattern of recovery of the affected biocoenoses in the next two years. Results show an impressive and fast shift of the benthic assemblage from a biocoenosis mainly composed of slow-growing and long-lived species to a biocoenosis dominated by fast-growing and short-lived species. The sponge Chondrosia reniformis, one of the key species of this assemblage, which had never been involved in previous massive mortality events in the Mediterranean Sea, reduced its coverage by 70%, and only few small specimens survived. All the damaged sponges, together with many associated organisms, were detached by rough-sea conditions, leaving large bare areas on the rocky wall. Almost three years after the disease, the survived specimens of C. reniformis did not increase significantly in size, while the bare areas were colonized by fast-growing species such as stoloniferans, hydrozoans, mussels, algae, serpulids and bryozoans. Cnidarians were more resilient than massive sponges since they quickly recovered in less than one month. In the study area, the last two outbreaks caused a reduction in the filtration efficiency of the local benthic assemblage by over 60%. The analysis of the times series of wave heights and temperature revealed that the conditions in summer 2011 were not so extreme as to justify severe mass mortality, suggesting the occurrence of other factors which triggered the disease. The long-term observations of a benthic assemblage in the NW Adriatic Sea allowed us to monitor its dynamics before, during and after the mortality event. The N Adriatic Sea responds quickly to climatic anomalies and other environmental stresses because of the reduced dimension of the basin. The long-term consequences of frequent mass mortality episodes in this area could promote the shift from biocoenoses dominated by slow-growing and long-lived species to assemblages dominated by plastic and short life cycle species.

Growth of the tropical zoanthid Palythoa caribaeorum (Cnidaria: Anthozoa) on reefs in northeastern Brazil.

In Brazilian reefs, zoanthids, especially Palythoa caribaeorum are fundamental for structuring the local benthic community. The objective of this study was to determine the growth rate of P. caribaeorum, and to assess the influence of the site (different beaches), season (dry and wet), location (intertidal or infralittoral zones), and human pressure associated with tourism. For one year we monitored the cover of P. caribaeorum in transects and focused on 20 colonies. We cut off a square (100 cm2) from the central part of the colony and monitored the bare area for four months in each season. The average growth rates varied from 0.015 and 0.021 cm.day(-1). The rate was homogeneous in all localities, and there was no influence from colony site, location, or touristic visitation, showing that the growth velocity may be an intrinsic characteristic of the species, with a strong genetic component. The growth rate of P. caribaeorum differed among months, and peaked in the first month after injury. The average cover varied from 6.2 to 22.9% and was lower on the reef visited by tourists. The present study corroborates the hypothesis that P. caribaeorum is important for coastal reef dynamics due to its fast and continuous growth.

Larval nervous systems: true larval and precocious adult.

The apical organ of ciliated larvae of cnidarians and bilaterians is a true larval organ that disappears before or at metamorphosis. It appears to be sensory, probably involved in metamorphosis, but knowledge is scant. The ciliated protostome larvae show ganglia/nerve cords that are retained as the adult central nervous system (CNS). Two structures can be recognized, viz. a pair of cerebral ganglia, which form the major part of the adult brain, and a blastoporal (circumblastoporal) nerve cord, which becomes differentiated into a perioral loop, paired or secondarily fused ventral nerve cords and a small perianal loop. The anterior loop becomes part of the brain. This has been well documented through cell-lineage studies in a number of spiralians, and homologies with similar structures in the ecdysozoans are strongly indicated. The deuterostomes are generally difficult to interpret, and the nervous systems of echinoderms and enteropneusts appear completely enigmatic. The ontogeny of the chordate CNS can perhaps be interpreted as a variation of the ontogeny of the blastoporal nerve cord of the protostomes, and this is strongly supported by patterns of gene expression. The presence of 'deuterostomian' blastopore fates both in an annelid and in a mollusk, which are both placed in families with the 'normal' spiralian gastrulation type, and in the chaetognaths demonstrates that the chordate type of gastrulation could easily have evolved from the spiralian type. This indicates that the latest common ancestor of the deuterostomes was very similar to the latest common pelago-benthic ancestor of the protostomes as described by the trochaea theory, and that the neural tube of the chordates is morphologically ventral.

How do environmental factors influence life cycles and development? An experimental framework for early-diverging metazoans.

Ecological developmental biology (eco-devo) explores the mechanistic relationships between the processes of individual development and environmental factors. Recent studies imply that some of these relationships have deep evolutionary origins, and may even pre-date the divergences of the simplest extant animals, including cnidarians and sponges. Development of these early diverging metazoans is often sensitive to environmental factors, and these interactions occur in the context of conserved signaling pathways and mechanisms of tissue homeostasis whose detailed molecular logic remain elusive. Efficient methods for transgenesis in cnidarians together with the ease of experimental manipulation in cnidarians and sponges make them ideal models for understanding causal relationships between environmental factors and developmental mechanisms. Here, we identify major questions at the interface between animal evolution and development and outline a road map for research aimed at identifying the mechanisms that link environmental factors to developmental mechanisms in early diverging metazoans. Also watch the Video Abstract.

Lineage-specific evolution of cnidarian Wnt ligands.

We have studied the evolution of Wnt genes in cnidarians and the expression pattern of all Wnt ligands in the hydrozoan Hydractinia echinata. Current views favor a scenario in which 12 Wnt sub-families were jointly inherited by cnidarians and bilaterians from their last common ancestor. Our phylogenetic analyses clustered all medusozoan genes in distinct, well-supported clades, but many orthologous relationships between medusozoan Wnts and anthozoan and bilaterian Wnt genes were poorly supported. Only seven anthozoan genes, Wnt2, Wnt4, Wnt5, Wnt6, Wnt 10, Wnt11, and Wnt16 were recovered with strong support with bilaterian genes and of those, only the Wnt2, Wnt5, Wnt11, and Wnt16 clades also included medusozoan genes. Although medusozoan Wnt8 genes clustered with anthozoan and bilaterian genes, this was not well supported. In situ hybridization studies revealed poor conservation of expression patterns of putative Wnt orthologs within Cnidaria. In polyps, only Wnt1, Wnt3, and Wnt7 were expressed at the same position in the studied cnidarian models Hydra, Hydractinia, and Nematostella. Different expression patterns are consistent with divergent functions. Our data do not fully support previous assertions regarding Wnt gene homology, and suggest a more complex history of Wnt family genes than previously suggested. This includes high rates of sequence divergence and lineage-specific duplications of Wnt genes within medusozoans, followed by functional divergence over evolutionary time scales.

The invasive snowflake coral (Carijoa riisei) in the Tropical Eastern Pacific, Colombia / El coral copo de nieve invasivo (Carijoa riisei) en el Pacífico Oriental Tropical, Colombia

Carijoa riisei (Octocorallia: Cnidaria), a western Atlantic species, has been reported in the Pacific as an invasive species for nearly forty years. C. riisei has been recently observed overgrowing native octocorals at several rocky-coral littorals in the Colombian Tropical Eastern Pacific-(TEP). C. riisei has inhabited these reefs for at least 15 years but the aggressive overgrowth on other octocorals have been noted until recently. Here, we surveyed for the first time the distribution and inter-specific aggression by C. riisei in both coastal and oceanic areas colonized in the Colombian TEP (Malpelo, Gorgona and Cabo Corrientes), including preliminary multiyear surveys during 2007-2013. We observed community-wide octocoral mortalities (including local extinction of some Muricea spp.) and a steady occurrence of competing and overgrowing Pacifigorgia seafans and Leptogorgia seawhips. In Gorgona Island, at two different sites, over 87% (n=77 tagged colonies) of octocorals (Pacifigorgia spp. and Leptogorgia alba) died as a result of C. riisei interaction and/or overgrowth between 2011 and 2013. C. riisei overgrows octocorals with an estimate at linear growth rate of about 1cm m-1. The aggressive overgrowth of this species in TEP deserves more attention and regular monitoring programs. Rev. Biol. Trop. 62 (Suppl. 1): 199-207. Epub 2014 February 01.

Carijoa riisei (Octocorallia: Cnidaria), una especie nativa del Atlántico Occidental. Sin embargo, se ha reportado como especie exótica invasora en el Pacífico desde hace unos cuarenta años y en la costa occidental de Colombia hace 15-20 años. Recientemente C. riisei se ha observado sobrecreciendo octocorales nativos en varios litorales rocosos coralinos en el Pacífico Oriental Tropical-(POT) de Colombia. C. riisei ha habitado estos arrecifes durante al menos 15 años, pero este comportamiento agresivo frente a otros octocorales no se habían notado hasta hace unos pocos años. En este estudio, se evalúa por primera vez, en las zonas costeras y oceánicas, la distribución e incidencia C. riisei en otros octocorales del POT colombiano (Malpelo, Gorgona y Cabo Corrientes), incluyendo información observaciones entre 2007-2013. Hemos observado mortalidad en toda la comunidad de octocorales (incluyendo la extinción local de algunas especies de Muricea) y una constante competencia y cubrimiento de abanicos de mar Pacifigorgia y gorgonáceos Leptogorgia. C. riisei presento una tasa lineal de crecimiento sobre otros octocorales de aproximadamente 1 cm m-1. El comportamiento agresivo de C. riisei en el POT merece más atención y programas regulares de monitoreo.

Aryl hydrocarbon receptor (AHR) in the cnidarian Nematostella vectensis: comparative expression, protein interactions, and ligand binding.

The aryl hydrocarbon receptor (AHR) is a member of the basic helix-loop-helix/Per-ARNT-Sim (bHLH-PAS) family of transcription factors and has diverse roles in development, physiology, and environmental sensing in bilaterian animals. Studying the expression of conserved genes and function of proteins in outgroups to protostomes and deuterostomes assists in understanding the antiquity of gene function and deciphering lineage-specific differences in these bilaterian clades. We describe the developmental expression of AHR from the sea anemone Nematostella vectensis and compare its expression with three other members of the bHLH-PAS family (AHR nuclear translocator (ARNT), Cycle, and a proto-Single-Minded/Trachealess). NvAHR expression was highest early in the larval stage with spatial expression in the basal portion of the ectoderm that became increasingly restricted to the oral pole with concentrated expression in tentacles of the juvenile polyp. The other bHLH-PAS genes showed a divergent expression pattern in later larval stages and polyps, in which gene expression was concentrated in the aboral end, with broader expression in the endoderm later in development. In co-immunoprecipitation assays, we found no evidence for heterodimerization of AHR with ARNT, contrary to the conservation of this specific interaction in all bilaterians studied to date. Similar to results with other invertebrate AHRs but in contrast to vertebrate AHRs, NvAHR failed to bind two prototypical xenobiotic AHR ligands (2,3,7,8-tetrachlorodibenzo-p-dioxin, ß-naphthoflavone). Together, our data suggest that AHR's original function in Eumetazoa likely involved developmental patterning, potentially of neural tissue. The role of heterodimerization in the function of AHR may have arisen after the cnidarian-bilaterian ancestor. The absence of xenobiotic binding to NvAHR further supports a hypothesis for a derived role of this protein in chemical sensing within the chordates.

Expression of Wnt pathway genes in polyps and medusa-like structures of Ectopleura larynx (Cnidaria: Hydrozoa).

The canonical Wnt signaling pathway is conserved in its role in axial patterning throughout Metazoa. In some hydrozoans (Phylum Cnidaria), Wnt signaling is implicated in oral-aboral patterning of the different life cycle stages-the planula, polyp and medusa. Unlike most hydrozoans, members of Aplanulata lack a planula larva and the polyp instead develops directly from a brooded or encysted embryo. The Aplanulata species Ectopleura larynx broods such embryos within gonophores. These gonophores are truncated medusae that remain attached to the polyps from which they bud, and retain evolutionary remnants of medusa structures. In E. larynx, gonophores differ between males and females in their degree of medusa truncation, making them an ideal system for examining truncated medusa development. Using next-generation sequencing, we isolated genes from Wnt signaling pathways and examined their expression in E. larynx. Our data are consistent with the Wnt pathway being involved in axial patterning of the polyp and truncated medusa. Changes in the spatial expression of Wnt pathway genes are correlated with the development of different oral structures in male and female gonophores. The absence of expression of components of the Wnt pathway and presence of a Wnt pathway antagonist SFRP in the developing anterior end of the gonophore suggest that downregulation of the Wnt pathway could play a role in medusa reduction in E. larynx.